The present disclosure relates to an image obtaining apparatus, an image obtaining method, and an image obtaining program obtaining an image by using a microscope.
Cancer cells of some breast-cancer patients have more HER-2 receptors, a kind of protein, than normal cells do. In view of this, a surgically-resected sample is examined by the HER-2 (Human Epithelialgrowth factor Receptor type 2) examination method. Based on the result, a medicine, which will be administered to a patient after the operation, is selected. In the HER-2 examination method, a surgically-resected sample (mammary tissue) is stained with a reagent called PathVysion in HER-2 DNA probe kit. The reagent includes probes. One probe hybridizes HER-2/neu genes, which code HER-2 proteins. The other probe hybridizes alpha-satellite DNA sequences in a centromere area of chromosome 17.
When a sample is irradiated with an excitation light, which excites the probes, the probes are excited and emit fluorescence. In this case, the wavelength of the fluorescence emitted from the probe, which hybridizes HER-2/neu genes, is different from the wavelength of the fluorescence emitted from the probe, which hybridizes alpha-satellite DNA sequences. In other words, HER-2/neu genes emit red fluorescence, and alpha-satellite DNA sequences emit green fluorescence. In diagnosis, it is determined that HER-2/neu genes are increased or not based on the ratio between the number of red bright points and the number of green bright points. The red bright points are fluorescent labels showing HER-2/neu genes in a cell nucleus. The green bright points are fluorescent labels showing alpha-satellite DNA sequences in a cell nucleus. In a case where the ratio of the number of the fluorescent labels showing HER-2/neu genes to the number of the fluorescent labels showing alpha-satellite DNA sequences is equal to or more than 2.2, HER-2 positive reaction is determined (see HER-2 Examination Guide, 3rd edition, Trastuzumab Pathology Working Group, September 2009, p. 10 <FISH-method determination method>).
Further, Japanese Patent Application Laid-open No. 2011-107669 discloses a technology of detecting bright points, which mark a target cell, from a fluorescent image of a biological sample. Japanese Patent Application Laid-open No. 2011-107669 discloses a biological sample image obtaining apparatus. The biological sample image obtaining apparatus moves the focus of an objective lens in the thickness direction of a target site of a biological sample. At the same time, the biological sample image obtaining apparatus moves the image of a site magnified by the objective lens, which is formed by an image sensor, in the surface direction. The biological sample image obtaining apparatus is capable of distinguishing bright points, which are generated by fluorescent markers emitting light, from bright points, which are generated by noises. As a result, accuracy of detecting bright points may be improved.